BULLETIN OF THE ALLYN MUSEUM Published By The McGuire Center for Lepidoptera and Biodiversity Florida Museum of Natural History University of Florida P. O. Box 112710 Gainesville, FL 32611-2710 Number 166 5 November 2012 ISSN-0097-3211 A NEW SPECIES OF OPSIPHANES DOUBLEDAY, [1849] FROM COSTA RICA (NYMPHALIDAE: MORPHINAE: BRASSOLINI), AS REVEALED BY ITS DNA BARCODES AND HABITUS Isidro A. Chacón1, José Montero-Ramírez1, Daniel H. Janzen2, Winnie Hallwachs2, Patrick Blandin3, C. R. Bristow4, and Mehrdad Hajibabaei5 1Instituto Nacional de Biodiversidad (INBio), Apdo. 22-3100, Santo Domingo, Heredia, Costa Rica, e-mails: [email protected] and [email protected] 2Department of Biology, University of Pennsylvania, Philadelphia, PA 19104, USA e-mails: [email protected] and [email protected] 3Entomologie, Muséum national d’Histoire naturelle, 57, rue Cuvier, 75005 Paris, France e-mail: [email protected] 42 Sid Bank, Sid Lane, Sidmouth Devon EX10 5NX UK, e-mail [email protected] 5Biodiversity Institute of Ontario, Department of Integrative Biology, University of Guelph, Guelph, Ontario, Canada N1G 2W1 ABSTRACT: Opsiphanes jacobsorum new species (Nymphalidae) is described from Costa Rica with descriptions of habitat, host plants, last instar larva and pupa. The holotype is DNA barcoded (COI), and a cladogram of the species of Opsiphanes present in Area Conservacion Guanacaste (ACG) is included. We review this new species in the context of the Costa Rican species of Opsiphanes and specifically the complex of Opsiphanes cassina C. Felder & R. Felder, 1862. KEY WORDS: Area de Conservacion Guanacaste, Brassolini, caterpillar inventory, Costa Rica, Lepidoptera, new species, Nymphalidae, Opsiphanes, palm-eating caterpillars, rain forest, rare species. INTRODUCTION In the ongoing caterpillar inventory of the northwest Costa Rican low to middle- elevation forests of Area de Conservacion Guanacaste (ACG) (Janzen et al. 2009, 2011), the large caterpillars of five medium-sized species of Brassoline butterflies Opsiphanes quiteria (Stoll,1780), O. tamarindi C. Felder & R. Felder, 1861, O. bogotanus Distant,1875, 2 O. invirae (Hübner, [1808]), and O. fabricii (Boisduval, 1870) have been encountered more than 3,300 times in the past 34 years. The caterpillars of a much rarer sixth species, O. zelotes Hewitson, 1873, have also been encountered 16 times (Janzen and Hallwachs 2012), though in contrast to the former five species, free-flying adults of O. zelotes have not been encountered in nature in ACG. However, during the intense caterpillar inventory of the mid-elevation rain forest (400-600 m) in Sector Pitilla of northeastern ACG, a seventh species of Opsiphanes has been encountered at very low density. The male adults are very similar to the males of the common O. fabricii widely called O. cassina, in Costa Rica (see DeVries (1987)), and designated as O. cassina fabricii by DeVries (1987) and Bristow (1991), but separated from the nominate O. cassina by Blandin (1977). The male of this seventh species was initially noticed because it has different DNA barcodes but then it was realized that its color pattern is slightly outside the range of variation of O. fabricii males in ACG. Upon a close scrutiny, its female is easily differentiated from its larger look-alike, the female of the Central American subspecies of O. quiteria quirinus. Thus, considering the present state of knowledge on the genus Opsiphanes (Bristow 1991; Casagrande 2004), this ACG species is new. Here we describe this species as a new member of the well-known Costa Rican, and hence Central American, butterfly fauna, and hope it will no longer be overlooked amongst other similar species of Opsiphanes. Its cryptic presence also suggests the usefulness of close examination of any seemingly variable single species of Opsiphanes, so as to determine to what degree it might be a complexes of cryptic species (as has proven to be the case with numerous other ACG species of large and conspicuous Lepidoptera (Janzen et al. 2009, 2011)). MATERIALS AND METHODS Genitalia (Figs. 9-14) dissections and measurements were made with an Olympus SZ60 stereomicroscope with a calibrated ocular micrometer. The following protocol was used for the dissection of genitalia. Abdomens were prepared using a 10% solution of KOH. Mercurochrome and Eosin Y were used as stains. The genitalia and pelt were subsequently stored in glycerol for examination (70% ethanol solution, 3:1). The genitalia and the pelt were slide mounted using Euparal. Genitalic slides were photographed using a JVC 3-CCD color video camera through an Olympus SZ60 stereomicroscope, both mounted in an Olympus SZH-ILLD illumination base and by using Montage explorer software (version: 2.01.0075, Synoptics Ltd.) and Auto-Montage software (version: 4.02.0014, Synoptics Ltd.). Morphological terminology follows Klots (1970) and Kristensen (2003). Acronyms for depositories of specimens are as follows: INBio - Instituto Nacional de Biodiversidad, Santo Domingo de Heredia, Costa Rica; MNHN – Muséum national d’Histoire naturelle, Paris, France; FLMNH/MGCL – McGuire Center for Lepidoptera and Biodiversity; USNM – National Museum of Natural History, Washington D.C, USA. All images here and many others for comparison are available on the ACG caterpillar inventory web site (Janzen and Hallwachs 2012). Taxonomic Background and Methodological Considerations This new species description is constructed on the fundamental revision of Opsiphanes by Bristow (1991) and Blandin (1977). We take into account all the previously described species of Opsiphanes, which is important because all the previously known ACG species 3 belong to widely distributed classical species: Opsiphanes quiteria and O. invirae exist throughout the Neotropical Region, from Central America to southeastern Brazil; the complex O. cassina/fabricii (see below) is distributed from Mexico to western Ecuador, western and eastern Colombia, Venezuela and the Guianas, northeastern Brazil (Para), eastern Ecuador, Peru and Bolivia; O. tamarindi exists in Central America, Colombia, Ecuador (on both sides of the Andes) and may be in Peru (depending on taxonomists’ viewpoints); O. bogotanus also exists in Central America, Colombia, and western Ecuador; even the very rare O. zelotes is known from Central America and Colombia. Our new species is very similar to O. fabricii and it appears as its sister-species in the Neighbor Joining Tree (Fig. 27). Within the ACG fauna, O. “fabricii” is an obvious entity and was raised to species status by Blandin (1977). In concordance with Bristow (1991), we feel that the O. “fabricii” ACG population is O. cassina fabricii and therefore now to be called O. fabricii, and is distinct from O. cassina chiriquensis by both habitus and DNA barcode. However, there are nomenclatural problems within the cassina/fabricii complex, that need further review. Opsiphanes cassina was described in 1862 from western Amazonia (Bristow 1991). Its main diagnostic character is the existence of a limited, ochre-orange patch in the forewing cell, which however does not form a band crossing the cell as in the Central American taxon O. fabricii (1870), which has long been considered to be a subspecies of O. cassina (DeVries 1987; Bristow 1991). Later, the subspecies O. merianae Stichel, 1902 (Surinam), O. notanda Stichel, 1904 (Peru), O. numatius Fruhstorfer, 1912 (Colombia) = O. periphetes Fruhstorfer, 1912 (Colombia) were described. All of these species have an orange band across the forewing cell, as in O. fabricii. However, in a survey of the brassoline biogeography, Blandin (1977) noted that in the MNHN, there are an O. cassina male and an O. “fabricii” male from the same exact Ecuadorian locality. He proposed to separate O. cassina (reduced cell patch) from the O. “fabricii/notanda/etc.” group (with a complete cell band). This group is named O. fabricii according to taxonomic priority, and which we follow here. Bristow (1991) did not consider this point, and included O. fabricii, O. notanda, and other subspecies, including new ones, in O. cassina. There are now many specimens from northern Peru (San Martin, Amazonas) and Ecuador that confirm the sympatry between O. cassina and O. “fabricii”-like specimens (Blandin, paper in prep.). The ACG population belongs to the nominate O. fabricii fabricii as defined under O.cassina fabricii by Bristow (1991). Opsiphanes jacobsorum Chacón & Montero, new species (Figs.1-4, 9-14, 16, 18, 20, 21-22, 24, 26) Diagnosis. Among known species of Opsiphanes, the male of O. jacobsorum is similar only to O. fabricii and to O. cassina in that it has yellowish-orange in the forewing cell (see images in DeVries (1987), Bristow (1991)). It is close to O. fabricii, but it can be distinguished by the yellowish-orange patch in the forewing cell being widely separate from the forewing diagonal band rather than touching or even merging with it as is the case with O. fabricii and O. cassina. The female is superficially similar to the females of O. fabricii and of O. cassina except that the broad diagonal band across the forewing is white instead of yellow as in O. fabricii and O. cassina. Thus, the female looks like a small female of O. quiteria quirinus. Compare figures 1-4 with images of ACGO. fabricii (Fig. 5-8) and many more displayed at http://janzen.bio.upenn.edu/caterpillars/database.lasso). Description. Male – forewing length 34.70 mm (Figs. 1-2); venation typical for Opsiphanes (e. g., Stichel 1904). 4 Figures 1-4. Male holotype and female paratype of Opsiphanes jacobsorum: 1-2) male (06-SRNP- 34098), forewing length 34.70 mm; 3-4) female (06-SRNP-34095), forewing length 45.50 mm. Head: Orange-brown clubbed antennae, dark-brown between each flagellomere. Compound eyes black and rounded, surrounded by a ring of beige scales. Frons and vertex with a mix of beige and dark-brown piliform scales. Pointed labial palpi beige on dorsum, with a dark-brown line in the middle from base to apex. Dark, shiny black proboscis. Thorax: Dorsum dark-brown with a mix of beige and black scales; anterior part with beige and bright black, piliform scales, posterior part beige; tegulae dark brown.